The primary aim of this research is directed towards an understanding of cell wall biosynthesis on the molecular level. In directing our goals, a major emphasis will be placed on the role of the membrane in this process. Knowledge of the membrane-associated enzymes and carriers that are involved in the synthesis of this macromolecular net must be integrated with the control systems that govern the synthesis, assembly, and amplification of this structure. Seven specific projects will be investigated: (1) Mechanistic analysis of phospho-MurNAc-pentapeptide translocase - purification and properties of the enzyme. (2) Biosynthesis of spin-labeled peptidoglycan: analysis and interpretation of spectra, isolation and characterization of products. (3) Biosynthesis of membrane teichoic acid: role of the membrane in the incorporation of D-alanine into teichoic acid. (4) Mechanism of vancomycin action in the in vitro peptidoglycan system from Gaffkya homari. (5) Role of UDP-MurNAc-Ala-gamma DGlu-Lys-(epsilon-Ala) aDAla-DAla in the biosynthesis of cross-linked peptidoglycan. (6) Control of peptidoglycan biosynthesis in membrane vesicles: mechanism of action of effectors. (7) Characterization of nascent and released peptidoglycan in systems from E. coli and Gaffkya homari. An investigation of these projects will contribute to an understanding of cell biosynthesis and the role of the membrane in this process.